摘要
目的建立一种能够快速、灵敏、特异的鉴定结核分枝杆菌的实时荧光定量PCR(real-time quantitative polymerase chain reaction,qPCR)检测方法。方法根据结核分枝杆菌基因保守序列设计引物,并构建标准品。对确诊肺结核病人培养涂阳的结核痰液标本113份。正常人的痰液58份。戈登分枝杆菌、金黄色葡萄球菌、肺炎克雷伯球菌、表皮葡萄球菌、肺炎链球菌痰液标本共计30份。应用SYBR Green实时荧光定量PCR法、痰涂片抗酸染色法检测结核分枝杆菌。结果SYBRG Geen实时荧光定量PCR法、痰涂片抗酸染色法检测的特异性分别为99.1%、43.3%。结论荧光定量PCR是一种能够快速、灵敏度高、特异性强的结核分枝杆菌辅助诊断方法,具有重要的应用价值。
Objective:To develop a SYBR Green real-time fluorescent quantitative PCR system in identification of Mycobacterium tubercuLosis with characteristics of celerity,sensitivity and specificity.Methods:Detection of Mycobacterium tubercuLosisby SYBR Green real-time fluorescent quantitative PCR system.Evaluation of sensitivity,specificity and stability of this method.The primers were designed according to the conserved gene sequences of Mycobacterium tubercuLosis and the standard products were constructed.113 smear smear sputum specimens were collected from patients with confirmed tubercuLosis.58 sputum from normal people.A total of 30 sputum samples of Mycobacterium Gordon,Staphylococcus aureus,Klebsiella pneumoniae,Staphylococcus epidermidis and Streptococcus pneumoniae were collected.Mycobacterium tubercuLosis was detected by fluorescence quantitative PCR and sputum smear acid fast staining.ResuLts:The specificity of fluorescence quantitative PCR assay and sputum smear acid fast staining method were 99.1%and 43.3%respectively.Conclusion:SYBR Real-time fluorescent quantitative PCR system for the detection of Mycobacterium tubercuLosis with high sensitivity,high specificity and high stability,which is also fast and easy to operate.Therefore,this approach will be be a fast and effective method for the detection of Mycobacterium tubercuLosis.
出处
《中国食品工业》
2018年第12期66-71,共6页
China Food Industry
基金
兰州市科技计划项目(2016-3-107)
兰州市科技创新项目(2015-RO70).
关键词
结核分枝杆菌
实时荧光定量
PCR
Mycobacterium tubercuLosis
Real-time fluorescent quantitative
PCR